TWI714069B - Flow cell with integrated manifold - Google Patents

Abstract

In one example, a flow cell includes a plurality of inlet ports sized to receive a flow of reagent from one of a plurality of reagents into the flow cell. An outlet port of the flow cell is sized to pass each flow of reagent out of the flow cell. A flow channel of the flow cell is positioned between, and in fluid communication with, each inlet port and the outlet port. The flow channel includes a manifold section and a detection section. The manifold section has a plurality of manifold branches in fluid communication with a common line, wherein each branch is connected to one of each inlet port. The detection section is in fluid communication with the common line and the outlet port. The detection section is operable to perform a plurality of different chemical reactions between the plurality of reagents and analytes positioned in the detection section.

Description

Flow cell with integrated manifold

The present disclosure relates to flow cells with integrated manifolds.

Many instruments using microfluidic devices may include multiple reagent wells containing various reagents, where each reagent well is connected to a rotary selection valve. The rotary valve is aligned with each well channel in order to select any one of the reagents. Then use the common pipeline to transfer the selected reagent from the rotary valve to the inlet of the flow cell in a prescribed route. Analytes (such as DNA fragments, nucleic acid strands, etc.) can be positioned in the flow channel. Selected reagents can flow through the flow cell to perform various controlled chemical reactions on the analyte.

In order to minimize and in some cases even completely eliminate cross-contamination of reagents, each reagent used in a sequence of chemical reactions is often removed from the common pipeline (or flushing reagent) outside the flow cell by the next reagent (or flushing reagent) in the sequence. That is, the external common pipeline) and the flow cell are both flushed out to achieve a predetermined flushing efficiency.

However, the reagents utilized in this orderly chemical reaction can be very expensive. In addition, achieving this level of flushing efficiency in the flow channel of the flow cell often requires flushing a certain volume of reagent through the flow channel, which is many times the swept volume of the flow channel. For example, the flushing efficiency to achieve a predetermined concentration of a reagent located in the flow channel may involve flushing a volume of that reagent through the flow channel that is 5 to 10 times the sweep volume of the flow channel.

One of the reasons why such a high volume of the reagent and therefore such a high flush factor is involved is that the sweep volume of the external common line in the instrument is often high compared to the sweep volume of the flow channel . The sweep volume of the common pipeline outside the flow cell is often two or more times the sweep volume of the flow cell itself, both of which can be flushed in order to achieve the flushing efficiency involved in an orderly chemical reaction.

In addition, the flow path through the external common pipeline and the flow path through the flow channel of the flow cell are often not in the same plane. For example, the common line may include fittings, manifolds, layers, materials, or the like that cause sharp bends in the flow path (eg, at a right angle or greater) for connection to the flow cell and/or rotary valve. In addition, as an example, the reagent wells are often located at a different level in the instrument from the flow cell, and the external common pipeline can often be adjusted for this difference.

These level changes and sharp bends can contribute to areas of flow that are significantly slower than the reagent flow through most of the flow path (in this case, dead zones). The dead zone may be a slow-moving laminar, vortex, or vortex area that can trap reagents and make it difficult to flush out the reagents. In some cases, these dead zones may require a considerable amount of flushing reagent volume to flush out previously located reagents (eg, remaining reagents) that were trapped in those dead zones and left behind after previous chemical reactions. In addition, fittings and other mechanical connections between the common line and the rotary valve or between the common line and the flow cell may also contribute to additional dead zones, which may increase the volume of flushing reagents involved in achieving a certain flushing efficiency.

The present disclosure provides a device for reducing the volume of reagent flow (ie, total flushing volume) involved in flushing a flow cell and reaching a predetermined level of reagent concentration (ie flushing efficiency) in the flow channel of the flow cell relative to the prior art, and Examples of methods. More specifically, the present disclosure provides an example of a flow cell, where the flow channel has a detection section and a manifold section integrated therein. The detection section is the area in the flow channel where the chemical reaction between the analyte and various reagents is performed. The manifold section provides an internal common pipeline area for the reagent flow before entering the detection section.

The present disclosure provides examples in which the manifold section is small relative to the detection section to reduce the total flushing volume used to achieve a certain flushing efficiency. The present disclosure provides examples where the manifold section and the detection section are in the same plane or are planar to help reduce the dead zone of the reagent flow. In addition, the present disclosure provides an example of a manifold section having a reagent flow path junction formed only at an acute angle to additionally illustrate the reduction of the dead zone of the reagent flow.

A flow cell according to one or more aspects of the present disclosure includes a plurality of inlets that are sized to receive a reagent flow from one of the plurality of reagents into the flow cell. The outlet of the flow cell is sized to allow each reagent flow to flow out of the flow cell. The flow channel of the flow cell is located between each inlet and outlet and is in fluid communication with each inlet and outlet. The flow channel includes a manifold section and a detection section. The manifold section has a plurality of manifold branches in fluid communication with the common pipeline, where each branch is connected to one of each inlet. The detection section is in fluid communication with the common pipeline and the outlet. The detection section is operable to perform a number of different chemical reactions between the analyte and the various reagents located in the detection section.

The instrument according to one or more aspects of the present disclosure includes a plurality of reagent wells. Each reagent well is operable to contain one of a variety of reagents located therein. The multiple valves of the instrument are in fluid communication with one of each reagent well. Each valve is operable to control the flow of reagent from the reagent well in communication with the valve. The flow cell is located in the instrument. The flow cell includes a plurality of inlets, outlets, and flow channels between them. Each inlet is in fluid communication with one of each valve, and each inlet is sized to receive one of each reagent flow. The outlet is sized to allow each reagent flow to flow out of the flow cell. The flow channel is in fluid communication with each inlet and outlet. The flow channel includes a manifold section and a detection section. The manifold section has a plurality of manifold branches in fluid communication with the common pipeline, with each branch connected to the inlet. The detection section is in fluid communication with the common pipeline and the outlet. The detection section is operable to perform a number of different chemical reactions between the analyte and the various reagents located in the detection section.

A method according to one or more aspects of the present disclosure includes connecting a flow cell to an instrument. The flow cell includes a plurality of inlets, outlets, and flow channels in fluid communication therebetween. The flow channel includes a manifold section and a detection section. The first valve among the plurality of valves of the instrument is operated to select the first reagent among the plurality of reagents. Each reagent is positioned in the corresponding reagent well of the instrument. The first reagent is pumped through the first inlet of the plurality of inlets of the flow cell and through the flow channel. Perform a first chemical reaction between the analyte and the first reagent located in the detection section of the flow channel. After the first chemical reaction is completed, at least some of the first reagent will remain in the flow channel as the remaining reagent. The subsequent valve among the plurality of valves is operated to select the subsequent reagent among the plurality of reagents. Subsequent reagents are pumped through the subsequent ones of the plurality of inlets and through the flow channel to flush out the remaining reagent from the flow channel. The remaining reagent is flushed out, so that at least about 99.95% of the concentration of the subsequent reagent in the detection section is reached in the total flush volume of the subsequent reagent, which is equal to about 2.5 times or less of the sweep volume of the flow channel .

Certain examples will now be described to provide a comprehensive understanding of the principles of the structure, function, manufacture, and use of the methods, systems, and devices disclosed herein. One or more examples are shown in the drawings. Those skilled in the art will understand that the methods, systems, and devices specifically described herein and shown in the accompanying drawings are non-limiting examples, and the scope of the present disclosure is only limited by the scope of patent applications. Features shown or described with respect to one example can be combined with features of other examples. Such modifications and changes are stipulated to be included in the scope of the present disclosure.

The terms "substantially", "approximately", "approximately", "relatively" or other such similar terms that can be used throughout the present disclosure (including the scope of patent applications) are used to describe and consider, for example, changes in processing Small fluctuations caused by deviations from references or parameters. Such small fluctuations also include zero fluctuations from a reference or parameter. For example, they can mean less than or equal to ± 10%, such as less than or equal to ± 5%, such as less than or equal to ± 2%, such as less than or equal to ± 1%, such as less than or equal to ± 0.5%, such as less than or equal to ± 0.2 %, for example, less than or equal to ±0.1%, for example, less than or equal to ±0.05%.

The flushing efficiency as used herein is the concentration in percentage of the volume of the flushing reagent left in the area of the flow channel where the analyte is located after the flushing operation. Generally, depending on the parameters of the chemical reaction to be performed, the ideal flushing efficiency to be achieved ranges from 96% to 100% concentration of the flushing reagent in the flow channel.

The swept volume as used herein is the internal volume of the component in the flow path of the reagent. Therefore, the swept volume of the flow channel is the total internal volume of the flow channel of the flow cell. In addition, the flushing factor as used herein is the volume of reagent flushed through the part, which is expressed in units of the swept volume of the part. Therefore, the total flush volume is the sweep volume multiplied by the flush factor.

So, for example, if the flow channel requires 10 times its sweep volume of reagent to be flushed through the flow channel to achieve a predetermined flushing efficiency, the flushing factor of the reagent that achieves the flushing efficiency is 10 (or in units of sweep volume). 10). In addition, if the flow channel has a sweep volume of 5 microliters, then the total flushing volume that achieves the flushing efficiency is 50 microliters (ie 5 microliters (sweep volume) x 10 (flushing factor)).

Figures 1A-4 show various examples of flow cells according to aspects disclosed herein. Figures 5-6 show various examples of instruments according to aspects disclosed herein. Figure 7 shows various examples of methods according to aspects disclosed herein.

1A and 1B, a perspective view (FIG. 1A) and a front side view (FIG. 1B) of a flow cell 100 with flow channels 102 are depicted. According to aspects disclosed herein, the flow channel 102 includes a manifold section 104 and a detection section 106 (best seen in Figure 2). The manifold section 104 and the detection section 106 are integrally connected together in fluid communication within the flow cell 100.

The flow cell 100 of FIGS. 1A and 1B further includes a top layer 108 that defines a top surface 116 of the flow channel 102 and a bottom layer 110 that defines a bottom surface 118 of the flow channel 102. The middle layer 112 is located between the top layer 108 and the bottom layer 110. The intermediate layer 112 defines the geometry of the flow channel 102.

The top layer 108, the bottom layer 110, and the intermediate layer 112 may be composed of glass, silicon, polymer, or other materials that can meet the application requirements of any of the layers 108, 110, and 112. Examples of polymers that can be used in any of the three layers 108, 110, 112 are: polycarbonate, polymethylmethacrylate, polyimide, polyethylene terephthalate, poly Ester, cyclic olefin copolymer (COC) and cyclic olefin polymer (COP). COC and COP are examples of optically transparent polymers, which are often used in the top layer 108 and the bottom layer 110. The three layers 108, 110, 112 may be composed of the same material, or they may be composed of different materials.

The three layers 108, 110, 112 can be joined together with various adhesives such as pressure sensitive or heat activated adhesives. In addition, the layers 108, 110, 112 may be thermally bonded or laser welded.

The intermediate layer 112 is shown as a single layer in FIGS. 1A and 1B. However, the intermediate layer 112 may be a stacked layer that is bonded together to define the geometry of the flow channel 102. In addition, for this stacked layer, the manifold section 104 may be made to have a different height from the detection section 106. For example, the intermediate layer 112 may be composed of a stack of 6 layers, where the bottom three layers of the stack constitute a manifold section, and the entire stack of 6 layers constitute a detection section.

The flow channel 102 of the flow cell 100 includes a gap height 114. The gap height 114 is defined by the distance between the bottom surface 118 of the flow channel and the top surface 116 of the flow channel. As shown in FIGS. 1A and 1B, the gap height 114 is substantially constant throughout the flow channel 102. As an example, the gap height 114 in some flow channels 102 may be between about 10 microns and about 100 microns. For example, the gap height 114 may be about 10 microns, about 20 microns, about 50 microns, about 60 microns, or about 100 microns.

With reference to Figure 2, a cross-sectional view of the flow cell 100 of Figure IB taken along line 2-2 is depicted in accordance with aspects disclosed herein. The flow cell 100 includes a plurality of inlets 120, 122, 124, 126, 128, 130 (herein, 120-130) and at least one outlet 132, with the flow channel 102 in between.

Each inlet 120-130 is sized to receive one from a variety of reagents 146, 148, 150, 152, 154, 156 (here 146-156) (best seen in Figure 5) The reagent flow (or flow path) (the flow path is represented by arrows 134, 136, 138, 140, 142, 144 (herein 134-144)) is received into the flow cell 100. The outlet 132 is sized to allow each flow path 134-144 of the reagent to flow out of the flow cell 100.

The flow channel 102 is located between each inlet 120-130 and outlet 132 and is in fluid communication with each inlet 120-130 and outlet 132. The flow channel 102 includes a manifold section 104 and a detection section 106 that are integrally connected and in fluid communication with each other.

The manifold section 104 has a plurality of manifold branches 160, 162, 164, 166, 168, 170 (160-170 herein) in fluid communication with the common line 172. Each branch 160-170 is connected to one of each inlet 120-130, respectively. The detection section 106 is in fluid communication with the common pipeline 172 and the outlet 132. The detection section 106 is operable to perform a plurality of different chemical reactions between the analyte (not shown) located in the detection section 106 and the various reagents 146-156. The analyte can be DNA fragments, oligonucleotides, other nucleic acid strands, etc.

The bottom surface 118 of the flow channel 102 is actually the top surface of the bottom layer 110 of the flow cell 100. Nanowells (not shown) may be patterned in the bottom surface 118 to capture the analyte. Alternatively, the bottom surface 118 may be coated by surface treatment to capture the analyte. In addition, a combination of nanopores and surface treatments can be used to capture analytes.

The reagents 146-156 can be used to perform many various controlled chemical reactions on the analytes arranged in the detection section 106. For example, the flow paths 134-144 of each reagent 146-156 can carry an identifiable label (such as fluorescently labeled nucleotide molecules, etc.) that can be used to label the analyte. Thereafter, the excitation light can be radiated through the top layer 108 and onto the analyte, so that the fluorescent label marked on the analyte causes the emitted photons to fluoresce. The emitted photon photons can be detected by the detection module 266 (best seen in Figure 6) of the instrument 200 during the detection process. (Note that in this particular example, the detection module 266 is the imaging module used during the imaging process.) Then, the device circuitry within the instrument 200 can process and transmit the data signals derived from those detected photons. The data signal can then be analyzed to reveal the nature of the analyte.

Although the detection module 266 is shown in this example as an imaging module for detecting photons of light, other forms of detection modules and detection schemes can be used to detect other forms of detectable properties related to the analyte. For example, the detectable properties associated with the analyte may include charge, magnetic field, electrochemical properties, pH changes, and the like. In addition, the detection module 266 may include a sensing device without limitation, which may be embedded in the flow cell 100, installed in an instrument outside the flow cell 100, or installed in any combination of the two.

Referring to FIG. 3, an enlarged view of the manifold section 104 of FIG. 2 is depicted in accordance with aspects disclosed herein. Advantageously, the manifold section 104 has a certain volume and geometry, which, compared to pre-existing technologies, significantly reduces the flushing of the flow cell 100 and the reagent concentration that reaches a predetermined level in the flow channel 102 of the flow cell 100 (I.e. flushing efficiency) (best seen in the curves 180, 182 and 184 of Figure 4) the amount of reagent flow involved (i.e. total flushing volume).

One such example of a geometry that reduces the flushing factor to achieve the desired flushing efficiency is the manner in which the manifold branches 160-170 are connected to the common line 172. More specifically, the manifold branches 160-170 of the manifold section 104 are in fluid communication with the common pipeline 172 through a plurality of forked joints 174, which guide each flow path 134-144 or reagents through the common pipeline 172 and enter the test Within paragraph 106. In the implementation shown, the forked joint 174 forms an acute angle 176 between the branches 160-170, which contain the flow paths of the reagents 146-156 in the plurality of flow paths 134-144. In some implementations, the fork-shaped joints 174 may all be only acute angles 176 or only some of the fork-shaped joints 174 may form acute angles 176.

The common line 172 is shown in FIG. 3 as a single common line in fluid communication between the manifold branches 160-170 and the detection section 106. However, the common pipeline 172 may also be a plurality of common pipelines in fluid communication between the manifold branches 160-170 of the flow channel 102 and the detection section 106.

By forming the junction 174 into an acute angle 176 (that is, an angle less than 90 degrees), the amount of dead space flowing at each junction can be reduced compared with the related art. That is, the tendency of the flow paths 134-144 to form eddies, vortices, slow laminar flow areas, etc., is greatly reduced because there are few sharp bends around which the flow paths can flow. Because dead zones may be difficult to flush, the reduction of these dead zones also reduces the flushing factor involved in achieving a predetermined flushing efficiency.

Another example of a geometric shape that reduces the flushing factor to achieve the desired flushing efficiency is that the manifold section 104 and the detection section 106 of the flow channel 102 are substantially on the same plane or flat. Therefore, there is no discontinuity or level change in the flow channel 102 that can cause dead zones (for example, vortex, vortex, etc.).

The volume of the manifold section 104 also helps to reduce the flushing factor and improve the flushing efficiency, because the sweep volume of the manifold section 104 is smaller than the sweep volume of the detection section 106. More specifically, in some implementations, the manifold section 104 may have a sweep volume that is at least about 10 times smaller than the sweep volume of the detection section 106. Additionally, in some implementations, the manifold section 104 may have a sweep volume that is at least about 20, 50, or 100 times smaller than the sweep volume of the detection section 106. Due to the small sweep volume of the manifold section 104, there are fewer reagents that need to be flushed to minimize and in some cases even completely eliminate reagent cross-contamination.

The flow cell 100 includes a plurality of inlets 120-130, where each inlet 120-130 is sized to receive a flow path 134-144 from one of the plurality of reagents 146-156 into the flow cell 100. Because in some implementations, each inlet 120-130 can only receive one reagent 146-156, the reagent flow paths 134-144 can be kept separate when they are outside the flow cell, and there can be no exterior that may be contaminated by other reagents. Public pipeline. In other words, in the instrument 200 (best seen in FIGS. 5 and 6) that includes the flow cell 100, the manifold section 104 of the flow channel 102 may be the only common area in the instrument 200, in which The different flow paths 134-144 of the different reagents 146-156 are routed together before flowing into the detection section 106 of the flow channel 102.

This means that only the flow channel 102 of the flow cell 100 located in the instrument 200 may need to be flushed to reduce and in some cases completely eliminate reagent cross-contamination, because the reagents 146-156 have separate Flow paths 134-144. It also means that the flushing factor involved in reaching the predetermined flushing efficiency of the flow cell 100 may be the same as the flushing factor involved in the instrument 200 including the flow cell 100.

Referring to FIG. 4, various curves 180, 182, 184 of the relationship between flushing efficiency and flushing factor are depicted in accordance with aspects disclosed herein. The flushing efficiency as used herein is the concentration in percentage of the volume of the flushing reagent left in the area of the flow channel (such as, for example, the detection section) where the analyte is located after the flushing operation. The flushing factor as used herein is the volume of reagent flushed through the part, which is expressed in units of the swept volume of the part.

More specifically, FIG. 4 shows three curves 180, 182, and 184. The curve 180 is a graph of the relationship between the flushing efficiency of the flow channel 102 of the flow cell 100 and the flushing factor according to the aspects disclosed herein, where the gap height 114 is 100 microns and the flow rate of the flushing reagent is 1500 microliters per minute. The curve 182 is a graph of the relationship between the flushing efficiency of the flow channel 102 of the flow cell 100 and the flushing factor according to aspects disclosed herein, where the gap height 114 is 60 microns and the flushing flow rate of the flushing reagent is 1500 microliters per minute. The curve 184 is a graph of the relationship between the flushing efficiency of the flow channel 102 of the flow cell 100 and the flushing factor according to the aspects disclosed herein, where the gap height 114 is 60 microns and the flushing flow rate of the flushing reagent is 500 microliters per minute.

It can be seen from the curves 180, 182, and 184 that, in all cases, the flow channel 102 includes a sweep volume and geometry so that the flushing efficiency is used to achieve a flushing efficiency of at least about 99.95% percent concentration of the reagent located in the detection section 106 The factor is about 3 or less, for example about 2.5 or less, about 2.3 or less (in units of swept volume). In addition, the flushing factor of 2.3 can achieve a flushing efficiency of at least about 99.95%, such as at least about 99.96%, at least about 99.97%, at least about 99.98%, at least about 99.99%, at least about 99.995% or higher. In addition, a flushing factor of 2.5 can achieve a flushing efficiency of at least about 99.95%, such as at least about 99.96%, at least about 99.97%, at least about 99.98%, at least about 99.99%, at least about 99.995% or higher. In addition, a flushing factor of 3.0 can achieve a flushing efficiency of at least about 99.95%, such as at least about 99.96%, at least about 99.97%, at least about 99.98%, at least about 99.99%, at least about 99.995% or higher. In addition, a flushing factor of 2.0 can achieve a flushing efficiency of at least about 99%, such as at least about 99.1%, at least about 99.2%, at least about 99.3%, at least about 99.4%, at least about 99.5% or higher. In comparison, in many cases, the pre-existing flow channel may involve a flushing factor of 4 to 5 units of the sweep volume of the pre-existing flow channel to achieve a flushing efficiency of at least about 99.95%.

The low flushing factor (eg, 2.5 or less) to achieve such a high flushing efficiency (eg, 99.95 or greater) may be due to several characteristics of the flow cell 100. For example, the manifold section 104 and the detection section 106 are integral parts of the flow channel 102 in the flow cell 100 and are in the same plane or flat. In addition, as an example, each inlet 120-130 of the flow cell 100 may only receive one kind of reagent 146-156, so that the flow paths 134-144 of the reagent are not transmitted together in a prescribed route until the manifold section 104. Also as an example, the manifold branches 160-170 of the manifold section 104 may form an acute angle at the junction 174. Also as an example, the manifold section 104 has a sweep volume that is at least about 10 times smaller than the sweep volume of the detection section 106.

In addition, because there is an inlet 120-130 for each reagent 146-156, the reagents can be kept separate in the instrument 200 (best seen in Figures 5 and 6) that includes the flow cell 100. Therefore, the manifold section 104 of the flow channel 102 includes the only common area in the instrument 200 in which the different flow paths 134-144 of the different reagents 146-156 are pressed before flowing into the detection section 106 of the flow channel 102 The prescribed routes are transmitted together.

Therefore, the curves 180, 182, 184 of the flow cell 100 may remain substantially unchanged regardless of the type of fluid connection used by the instrument 200 to connect the reagents 146-156 to the flow cell 100. For example, the fluid connection between the reagent well and the flow cell 100 can be rigidly connected to a substantially straight and flush metal tube, or the connection can be bent to accommodate the difference between the reagent well and the flow cell 100. Horizontal pipe connection.

Referring to Fig. 5, an example of a schematic diagram of a cassette 202 and an instrument 200 is depicted, wherein the cassette 202 contains a flow cell 100 according to aspects disclosed herein. In this particular example, the instrument 200 is a cartridge-based sequencing instrument, wherein the cartridge 202 of the sequencing instrument 200 includes a flow cell 100 and various reagent processing components. In addition, the box 202 may be detachable from the instrument 200 as a module, and the flow cell 100 may or may not be detachable from the box 202.

However, the flow cell 100 and the reagent processing part need not be connected to the instrument 200 through the box 202 interface. More precisely, they may be independent components separately installed in the instrument 200. In addition, the reagent processing part may not be detachable from the instrument alone, but the flow cell 100 may be detachable from the instrument.

The box 202 of the instrument 200 includes a plurality of reagent wells 204, 206, 208, 210, 212, 214 (herein 204-214), wherein each reagent well is operable to contain a plurality of reagents 146, 148, 150, 152,154,156 reagents. A plurality of well channels 216, 218, 220, 222, 224, 226 (herein 216-226) extend from each corresponding reagent well 204-214 to the corresponding inlet 120-130 of the flow cell 100, wherein each inlet is connected to Only one reagent 146-156 is in fluid communication.

Reagents 146-156 can be any of several types or combinations of reagents, depending on the type and sequence of chemical reactions to be performed at the flow cell. For example, reagents 146-156 may have the following types: ● The reagent 146 may be a blending mixture, which is a mixture of chemical substances that incorporate fluorescently labeled nucleotides into the DNA strand. ● The reagent 148 may be a scan mix, which is a mixture of chemicals that stabilize the DNA strand during the detection process. ● The reagent 150 may be a cleave mix, which is a mixture of chemical substances that enzymatically cleave fluorescently labeled nucleotides from the DNA strand. • The reagent 152 may be the first washing buffer, which is a mixture of washing reagents that removes active reagents from the flow cell. • The reagent 154 may be a second washing buffer, which is another mixture of washing reagents that removes active reagents from the flow cell. ● The reagent 156 can be air.

The cassette also includes a plurality of valves 228, 230, 232, 234, 236, 238 (228-238 herein) located in the orifice channels 216-226. Each valve 228-238 is in fluid communication with one of each reagent well 204-214. Each valve 228-238 is operable to control the flow path 134, 136, 138, 140, 142, 144 of the reagent from the reagent orifice 204-214 in communication with the valve 228-238. In this particular example shown in Figure 5, the valve is a pinch valve. However, other types of valves, such as solenoid valves, ball valves, etc., can also be used. In the specific configuration of FIG. 5, the instrument 200 does not include a rotary valve, which will select various reagents 146-156, and merge the flow paths 134-144 of the reagents into the common pipeline before entering the flow cell 100.

The flow cell 100 is located within the instrument 200 and may or may not be detachable from the cassette 202. In addition, if the cartridge 202 is not used, the flow cell 100 can also be detachable from the instrument 200.

The flow cell 100 includes a plurality of inlets 120, 122, 124, 126, 128, 130 and outlets 132. Each inlet 120-130 is in fluid communication with a corresponding valve 228-238 through a respective orifice channel 216-226. Each inlet 120-130 is sized to receive one of each flow path 134-144 of the reagent, respectively. The orifice channels 216-226 can be in various configurations. For example, the orifice channels 216-226 may be primarily metal tubes that rigidly connect the reagent orifices 204-214 to the inlets 120-130. Alternatively, the orifice channels 216-226 may be plastic tubes connecting the reagent orifices 204-214 to the inlets 120-130. The outlet 132 of the flow cell 100 is sized to allow each flow path 134-144 of the reagent to flow out of the flow cell 100.

The flow cell 100 includes a flow channel 102 located between each inlet 120-130 and outlet 132 and in fluid communication with each inlet 120-130 and outlet 132. The flow channel 102 includes a manifold section 104 and a detection section 106.

The manifold section 104 has a plurality of manifold branches 160, 162, 164, 166, 168, 170 in fluid communication with the common line 172. Each manifold branch 160-170 is connected to an inlet 120-130.

The detection section 106 is in fluid communication with the common pipeline 172 and the outlet 132. The detection section 106 is operable to perform a number of different chemical reactions between the analyte located in the detection section 106 and the various reagents 146-156.

The reagent flow paths 134-144 remain separated from each other until they enter the flow cell 100. Therefore, the manifold section 104 of the flow channel 102 includes the only common area in the instrument 200 where the different flow paths 134-144 of the different reagents 146-156 are combined before flowing into the detection section 106 of the flow channel 102. Transmission according to the prescribed route. Therefore, only the flow channel 102 needs to be flushed in order to minimize and in some cases even completely eliminate cross-contamination of reagents between chemical reactions. This helps to reduce the flushing factor and therefore the total flushing volume of the flushing reagent used to reach the predetermined concentration (rinsing efficiency) of the flushing reagent in the flow channel 102.

In addition, outside the flow cell 100 where the reagent flow paths 134-144 are kept separate, the reagent flow paths may flow more than one level. For example, the reagent wells 204-214 may be positioned in the instrument 200 at a higher level than the level of the flow cell 100. However, inside the flow cell 100 where the reagent flow paths 134-144 can be mixed, the manifold section 104 and the detection section 106 of the flow channel 102 are substantially on the same plane or flat. This helps reduce potential dead zones in the flow channel 102, and therefore also helps reduce the flushing factor used to achieve a predetermined flushing efficiency.

The outlet 132 of the flow cell 100 is in fluid communication with the first pump pinch valve 240. The first pump pinch valve 240 is in fluid communication with the second pump pinch valve 242.

An onboard pump 244 (for example, a syringe pump or the like) is also arranged on the cassette 202. Even though the onboard pump 244 may be another type of pump, it will also be referred to as a syringe pump 244 here. The syringe pump 244 is connected between the first pump pinch valve 240 and the second pump pinch valve 242 in a tee formation. Both the first pump pinch valve 240 and the second pump pinch valve 242 are opened and closed by the instrument 200 to enable the syringe pump 244 to be engaged with or disengaged from the flow cell 100.

The injection pump 244 includes a reciprocating plunger 246 arranged in a cylinder 248 having a cylinder bore 250. The plunger 246 is received in the cylinder bore 250 to form a plunger-cylinder bore seal. The plunger 246 is driven by the instrument 200 to reciprocate in the cylinder bore 250 and pump reagents from the reagent holes 204-214 to the waste tank 252.

Referring to FIG. 6, an example of a schematic block diagram of an instrument 200 including the detachable box 202 of FIG. 5 is depicted in accordance with aspects disclosed herein. The instrument 200 includes a docking station 260 for receiving the box 202. Various electrical and mechanical elements within the instrument 200 interact with the cassette 202 to operate the cassette 202 during a sequencing operation performed by the instrument 200.

The instrument 200 may include one or more processors 262 among other things. The processors 262 execute program instructions stored in the memory 264 to perform sequencing operations. The processor 262 electronically communicates with the detection module 266, the syringe pump drive element 268, the pinch valve drive element 270, and so on.

A user interface 274 is provided for the user to control and monitor the operation of the instrument 200. The communication interface 272 transmits data and other information between the instrument 200 and a remote computer, network, etc.

The syringe pump drive assembly 268 includes a syringe pump motor 276 coupled to the extendable shaft 278. The extendable shaft 278 is driven by the syringe pump motor 276 between the extended position and the retracted position to reciprocate the plunger 246 in the cylinder bore 250 of the cylinder 248 on the syringe pump 244.

The pinch valve drive assembly 270 includes a set of eight pneumatically driven pinch valve drive motors 280. Six of the pinch valve drive motors 280 are mechanically coupled to the pinch valves 228-238. Two of the pinch valve drive motors are mechanically coupled to the first pump pinch valve 240 and the second pump pinch valve 242. The pinch valve driving motor 280 can use air pressure to clamp or release the elastic center portions of the pinch valves 228-238, 240, 242 to open and close the pinch valves pneumatically. Alternatively, the pinch valve drive motor 280 may be electrically driven.

The detection module 266 includes all cameras and light detection sensors to detect the emitted light photons emitted from the analyte in the flow cell 100. The device circuitry (not shown) within the instrument 200 can then process and transmit the data signals derived from those detected photons. The data signal can then be analyzed to reveal the nature of the analyte.

Referring to FIG. 7, an example of a method of performing a series of experiments using the flow cell 100 is depicted according to aspects disclosed herein. This method utilizes an instrument 200 with a flow cell 100. The instrument 200 includes a plurality of reagent wells 204-214 containing a plurality of reagents 146-156. Each reagent well 204-214 is in fluid communication with a single inlet of the plurality of inlets 120-130 on the flow cell 100 so that the flow paths 134-144 of the reagents are not mixed until they enter the flow cell 100. The flow cell 100 includes a flow channel 102 having a manifold section 104 integrally connected to a detection section 106. The manifold section 104 receives the reagents 146-156 and transmits them through the common pipeline 172 to the detection section 106 according to a prescribed route. The analyte is located in the detection section 106, where multiple chemical reactions between the analyte and the reagents 146-156 are performed. Due to the geometry of the instrument 200 and the flow cell 100, the flushing factor (in units of sweep volume) is reduced compared to the flow cell of the prior art, and therefore used to flush the flow cell 100 and reach the flow channel 102 of the flow cell 100 The amount of reagent flow (that is, the total washing volume) for a predetermined level of reagent concentration (that is, washing efficiency).

The method (at step 300) begins by connecting the flow cell 100 to the instrument 200. The flow cell 100 includes a plurality of inlets 120-130, an outlet 132, and a flow channel 102 in fluid communication therebetween. The flow channel 102 includes a manifold section 104 and a detection section 106. Some features of the geometry and architecture of the flow cell 100 and the instrument 200 to reduce the flushing factor and improve the flushing efficiency are as follows: ● The manifold section 104 and the detection section can be on the same plane. ● The sweep volume of the manifold section 104 can be at least about 10 times smaller than the sweep volume of the detection section 106. ● The manifold branches 160-170 in the manifold section 104 may form an acute angle at the junction 174. ● Each inlet 120-130 can receive a reagent 146-156 flowing through a reagent flow path 134-144.

The manifold section 104 of the flow channel 102 may include the only common area in the instrument 200 in which the different flow paths 134-144 of the different reagents 146-156 are regulated before flowing into the detection section 106 of the flow channel 102 The routes are transferred together.

The method (at step 302) continues by operating a first valve of the plurality of valves 228-238 of the instrument 200 to select the first reagent of the plurality of reagents 146-156. Each reagent is positioned in the reagent wells 204-214 of the instrument 200.

The method (at step 304) continues by pumping the first reagent through the first of the plurality of inlets 120-130 and through the flow channel 102 of the flow cell 100. Pumping can be done with various suitable pumps. In the example shown in FIG. 5, the pump is a syringe pump 244.

The method (at step 306) continues by performing a first chemical reaction between the analyte located in the detection section 106 of the flow channel 102 and the first reagent. After the first chemical reaction is completed, a part of the first reagent will remain in the flow channel as the remaining reagent. The remaining reagents may have to be flushed out at least from the detection section 106 of the flow channel 102 in order to minimize and in some cases even completely eliminate the interaction with other reagents among the various reagents 146-156 used in the predetermined sequence of chemical reactions. Cross-contamination.

The method (at step 308) continues by operating a subsequent valve of the plurality of valves 228-238 to select the subsequent reagent of the plurality of reagents 146-156.

The method (at step 310) continues by pumping subsequent reagents through the subsequent ones of the plurality of inlets 120-130 and through the flow channel 102 to flush the remaining reagents from the flow channel 102 so that the subsequent ones located in the detection section 106 A concentration of at least about 99.95% of the reagent (ie, at least about 99.95% flushing efficiency) is achieved in the total flush volume of subsequent reagents, which is equal to about 2.5 times the sweep volume of the flow channel 102 (ie, about 2.5 The flushing factor) or less. This low flushing factor of 2.5 and a high flushing efficiency of 99.95% are achievable, at least in part due to the features discussed earlier at 300. Optionally (at step 310), for some subsequent reagents, the method may only use a flushing factor of about 2.0 or less to flush the remaining reagents from the flow channel 102 so that the subsequent reagents have a concentration of at least about 99% (ie, At least about 99% flushing efficiency) is achieved.

Then, the method (at step 312) continues by the following actions: after the concentration of at least about 99.95% (or alternatively at least about 99% for some subsequent reagents) of the subsequent reagents located in the detection section 106 is reached The subsequent chemical reaction between the subsequent reagent and the analyte located in the detection section 106. After the subsequent chemical reaction is completed, a part of the subsequent reagent will be left in the flow channel as the remaining reagent. This remaining reagent may have to be flushed out at least from the detection section 106 of the flow channel 102 in order to minimize and in some cases even completely eliminate other reagents among the various reagents 146-156 used in the chemical reaction of the predetermined sequence. Of cross contamination.

Then, the method (as shown by step 314) returns (to step 308) to repeatedly operate the subsequent valve (step 308), pump the subsequent reagents (step 310), and target multiple reagents in a predetermined sequence of chemical reactions. The predetermined sequence of reagents in 146-156 performs subsequent chemical reactions (step 312) to repeat the calculation.

With each iteration (step 314), the method can continue in a variety of ways according to the type of reagent selected. More specifically, the method can continue based on functional effects caused by chemical reactions between the analyte and various reagents.

For example, the method can be continued by incorporating fluorescently labeled nucleotides into an analyte that includes a DNA strand (ie, a DNA strand analyte). This can be achieved with blending mixtures such as reagent 146 or the like.

Also as an example, the method can be continued by stabilizing the DNA strand analyte during the detection process. This can be achieved by scanning a mixture such as reagent 148 or the like.

Also as an example, the method can be continued by enzymatically cleaving fluorescently labeled nucleotides from the DNA strand analyte. This can be achieved with a cleavage mixture such as reagent 150 or the like.

Before performing the chemical reaction between the reagent and the analyte positioned in the flow channel 102, not every flushing reagent requires a flushing efficiency of about 99.95% or greater (or alternatively, for some subsequent reagents, about 99%). % Or greater flushing efficiency). For example, if two washing buffer reagents are used sequentially, the second washing buffer may only require a washing efficiency of about 96% or more. In addition, for example, if the reagent is air that can be used to perform a predetermined in-situ test, the flushing efficiency may only be about 96%. However, in reagents of any given sequence, most subsequent reagents are more likely to require a flushing efficiency of 99.95% or greater (or alternatively, for some subsequent reagents, a flushing efficiency of about 99% or greater). This is especially the case when the subsequent reagent is not wash buffer or air. More specifically, this may be the case when the subsequent reagent is one of a blending mixture, a cleaving mixture, and a scanning mixture.

By making a flushing efficiency of at least about 99.95% in the case of a flushing factor of about 2.5 or less (or alternatively, at least about 99% in the case of a flushing factor of about 2.0 or less for some subsequent reagents) It is possible (especially for non-washing buffer reagents or non-air reagents). Compared with the prior art, the consumption of expensive reagents is reduced and the time to complete a series of controlled chemical reactions is significantly shortened. By making it possible to achieve a flushing efficiency of about 99.95% or greater with a flushing factor of about 2.5 or less (again, especially using non-wash buffer reagents or non-air reagents), the consumption of this expensive reagent The time elapsed to complete the sequence of controlled reactions is even further reduced and shortened even further.

It should be recognized that all combinations of the foregoing concepts and additional concepts discussed in more detail below (provided that such concepts are not inconsistent with each other) are contemplated as part of the inventive subject matter disclosed herein. In particular, all combinations of claimed subject matter appearing at the end of this disclosure are conceived as being part of the inventive subject matter disclosed herein.

Although the foregoing disclosure has been described with reference to specific examples, it should be understood that many changes can be made within the spirit and scope of the described creative concepts. Therefore, it is intended that the present disclosure is not limited to the described examples, but that it has a full scope defined by the language of the appended claims.

Various aspects of the present disclosure can be realized in one or more of the following embodiments:

1) A flow cell, including: A plurality of inlets, each of the plurality of inlets is manufactured according to a size to receive a reagent flow of a corresponding reagent from a plurality of reagents into the flow cell; An outlet, which is sized to allow the reagent flow to flow out of the flow cell; and A flow channel located between each of the plurality of inlets and the outlet and in fluid communication with each of the plurality of inlets and the outlet, the flow channel comprising: A manifold section having a plurality of manifold branches in fluid communication with a common pipeline, each of the plurality of manifold branches is connected to a corresponding one of the plurality of inlets, and The detection section is in fluid communication with the common pipeline and the outlet, and the detection section is operable to perform a plurality of different chemical reactions between the analyte located in the detection section and the plurality of reagents.

2) The flow cell according to 1), wherein the manifold section and the detection section of the flow channel are substantially planar.

3) The flow cell according to 1), wherein the plurality of manifold branches are in fluid communication with the common pipeline through a plurality of forked joints, and each of the plurality of forked joints guides a corresponding reagent The flow passes through the common pipeline and enters the detection section, and at least one fork-shaped joint forms an acute angle between the corresponding manifold branches.

4) The flow cell according to 1), wherein the manifold section has a sweep volume that is at least about 10 times smaller than the sweep volume of the detection section.

5) The flow cell according to 1), wherein the flow channel includes a sweep volume and a geometric shape, such that a flushing factor for achieving a flushing efficiency of at least about 99.95% concentration of the reagent located in the detection section is about 2.5 or less.

6) The flow cell according to 1), including: A top layer, which defines the top surface of the flow channel; A bottom layer, which defines the bottom surface of the flow channel; and An intermediate layer, which defines the geometry of the flow channel.

7) The flow cell according to 6), comprising a gap height defined by the distance between the bottom surface of the flow channel and the top surface of the flow channel, wherein the gap height is The flow channel is substantially constant and in the range of about 60 to 100 microns.

8) The flow cell according to 1), wherein the flow channel includes a sweep volume and a geometric shape such that the flushing factor for achieving a flushing efficiency of at least about 99% concentration of the reagent located in the detection section is About 2.0 or less.

9) The flow cell according to 1), wherein: The multiple reagents are from multiple corresponding reagent wells located in one of the cassette or the instrument, and One of the cartridge or the instrument includes a plurality of valves outside the flow cell, each valve being located between a corresponding reagent well of the plurality of corresponding reagent wells and a corresponding inlet of the plurality of inlets , So that each valve controls the reagent flow from the corresponding reagent well of the plurality of corresponding reagent wells.

10) An instrument including: A plurality of reagent wells, each of the plurality of reagent wells can be operated to contain reagents therein; A plurality of valves, each valve of the plurality of valves is in fluid communication with a corresponding reagent hole in the plurality of reagent holes, and each valve of the plurality of valves is operable to control the reagents from the plurality of reagents The reagent flow of the corresponding reagent well in the well; and A flow cell operable to be fluidly coupled to the instrument, the flow cell comprising: A plurality of inlets, each inlet being in fluid communication with a corresponding valve of the plurality of valves, and each inlet of the plurality of inlets is sized to receive the corresponding one from the plurality of reagent wells Reagent flow of reagent well; An outlet, which is sized to allow the reagent flow to flow out of the flow cell; and A flow channel located between each of the plurality of inlets and the outlet and in fluid communication with each of the plurality of inlets and the outlet, the flow channel comprising: A manifold section having a plurality of manifold branches in fluid communication with a common pipeline, each branch being connected to one of the plurality of inlets, and The detection section is in fluid communication with the common pipeline and the outlet, and the detection section is operable to perform a plurality of different chemical reactions between the analyte and various reagents located in the detection section.

11) The apparatus according to 10), wherein the manifold section of the flow channel is the only common area in which the flow path of the reagent is blocked before flowing into the detection section of the flow channel Transmitted together according to the prescribed route.

12) The instrument according to 10), wherein the manifold section and the detection section of the flow channel are substantially planar.

13) The apparatus according to 10), wherein the manifold branch is in fluid communication with the common pipeline through a plurality of fork-shaped joints, and the fork-shaped joints guide each reagent flow through the common pipeline and enter the common pipeline. In the detection section, the fork-shaped joint only forms an acute angle between the branches of the manifold.

14) The instrument according to 10), wherein the manifold section has a sweep volume that is at least about 10 times smaller than the sweep volume of the detection section.

15) The apparatus according to 10), wherein the flow channel includes a sweep volume and a geometric shape such that a flushing factor for achieving a flushing efficiency of at least about 99.95% concentration of the reagent located in the detection section is about 2.5 Or smaller.

16) The apparatus according to 12), including: A top layer, which defines the top surface of the flow channel; The bottom layer, which defines the bottom surface of the flow channel; An intermediate layer, which defines the geometry of the flow channel; and The gap height is defined by the distance between the bottom surface of the flow channel and the top surface of the flow channel, wherein the gap height is substantially constant throughout the flow channel.

17) The instrument according to 10), wherein the flow cell is positioned within the instrument.

18) A method including: Connecting a flow cell to the instrument, the flow cell including a plurality of inlets, outlets, and a flow channel in fluid communication between the plurality of inlets and the outlet, the flow channel including a manifold section and a detection section; Operate the first valve among the plurality of valves of the instrument to select the first reagent among the plurality of reagents, each of which is located in the cartridge or the reagent well of one of the instruments; Pumping the first reagent through the first inlet of the plurality of inlets of the flow cell and through the flow channel of the flow cell; Perform a first chemical reaction between the analyte located in the detection section of the flow channel and the first reagent, wherein after the first chemical reaction is completed, at least some of the first reagent Left in the flow channel as the remaining reagent; Operating a subsequent valve of the plurality of valves to select a subsequent reagent of the plurality of reagents; and Pump the subsequent reagents through the subsequent inlets of the plurality of inlets and through the flow channel to flush out the remaining reagents from the flow channel so that at least about 99.95% of the reagents located in the detection section The concentration of the reagent is the subsequent reagent using the total flush volume of the subsequent reagent, and the total flush volume is equal to or less than about 2.5 times the sweep volume of the flow channel.

19) The method according to 18), including: After the concentration of at least about 99.95% of the subsequent reagent located in the detection section is reached, a subsequent chemical reaction between the subsequent reagent located in the detection section and the analyte is performed, wherein After the subsequent chemical reaction is completed, at least some of the subsequent reagents are left in the flow channel as the second remaining reagent.

20) The method according to 19), comprising repeating the following actions: operating the subsequent valve, pumping the subsequent reagent, and performing a subsequent chemical reaction.

100‧‧‧Flow cell 102‧‧‧Flow Channel 104‧‧‧Manifold section 106‧‧‧Detection section 108‧‧‧Top floor 110‧‧‧Bottom 112‧‧‧Middle layer 114‧‧‧Gap height 116‧‧‧Top surface 118‧‧‧Bottom surface 120‧‧‧Entrance 122‧‧‧Entrance 124‧‧‧Entrance 126‧‧‧Entrance 128‧‧‧Entrance 130‧‧‧Entrance 132‧‧‧Exit 134‧‧‧Flow path 136‧‧‧Flow Path 138‧‧‧Flow Path 140‧‧‧Flow path 142‧‧‧Flow Path 144‧‧‧Flow Path 146‧‧‧Reagent 148‧‧‧Reagent 150‧‧‧Reagent 152‧‧‧Reagent 154‧‧‧Reagent 156‧‧‧Reagent 160‧‧‧Manifold branch 162‧‧‧Manifold branch 164‧‧‧Manifold branch 166‧‧‧Manifold branch 168‧‧‧Manifold branch 170‧‧‧Manifold branch 172‧‧‧Public pipeline 174‧‧‧Fork joint 176‧‧‧Acute Angle 180‧‧‧curve 182‧‧‧Curve 184‧‧‧Curve 200‧‧‧Instrument 202‧‧‧Box 204‧‧‧Reagent well 206‧‧‧Reagent well 208‧‧‧Reagent well 210‧‧‧Reagent well 212‧‧‧Reagent well 214‧‧‧Reagent well 216‧‧‧Hole Channel 218‧‧‧Hole Channel 220‧‧‧Hole Channel 222‧‧‧Hole Channel 224‧‧‧Hole Channel 226‧‧‧Hole Channel 228‧‧‧valve 230‧‧‧Valve 232‧‧‧valve 234‧‧‧valve 236‧‧‧valve 238‧‧‧valve 240‧‧‧Pinch valve for the first pump 242‧‧‧Pinch valve for second pump 244‧‧‧Board Pump/Syringe Pump 246‧‧‧Plunger 248‧‧‧Cylinder 250‧‧‧Cylinder bore 252‧‧‧ Waste Liquid Tank 260‧‧‧ docking station 262‧‧‧processor 264‧‧‧Memory 266‧‧‧Detection Module 268‧‧‧Syringe pump drive assembly 270‧‧‧Pinch valve drive assembly 272‧‧‧User Interface 274‧‧‧Communication interface 276‧‧‧Syringe pump motor 278‧‧‧Extendable shaft 280‧‧‧Pinch valve drive motor 300‧‧‧Step 302‧‧‧Step 304‧‧‧Step 306‧‧‧Step 308‧‧‧Step 310‧‧‧Step 312‧‧‧Step 314‧‧‧Step

According to the following detailed description in conjunction with the accompanying drawings, the present disclosure will be more fully understood. In the accompanying drawings: Figure 1A depicts an example of a perspective view of a flow cell with a flow channel according to aspects disclosed herein, wherein the flow channel includes a manifold section and a detection section; Figure 1B depicts an example of a front side view of the flow cell of Figure 1A according to aspects disclosed herein; Figure 2 depicts an example of a cross-sectional view of the flow cell of Figure IB taken along line 2-2 in accordance with aspects disclosed herein; Figure 3 depicts an example of an enlarged view of the manifold section of Figure 2 in accordance with aspects disclosed herein; Figure 4 depicts examples of various curves of the relationship between flushing efficiency and flushing factor according to aspects disclosed herein; FIG. 5 depicts an example of a schematic diagram of a cassette of an instrument containing the flow cell of FIG. 2 according to aspects disclosed herein; Figure 6 depicts an example of a schematic block diagram of an instrument including the cassette of Figure 5 according to aspects disclosed herein; and Figure 7 depicts an example of a flowchart of a method of performing a series of experiments using a flow cell according to aspects disclosed herein.

100‧‧‧Flow cell

102‧‧‧Flow Channel

104‧‧‧Manifold section

106‧‧‧Detection section

118‧‧‧Bottom surface

120‧‧‧Entrance

122‧‧‧Entrance

124‧‧‧Entrance

126‧‧‧Entrance

128‧‧‧Entrance

130‧‧‧Entrance

132‧‧‧Exit

134‧‧‧Flow path

136‧‧‧Flow Path

138‧‧‧Flow Path

140‧‧‧Flow path

142‧‧‧Flow Path

144‧‧‧Flow Path

160‧‧‧Manifold branch

162‧‧‧Manifold branch

164‧‧‧Manifold branch

166‧‧‧Manifold branch

168‧‧‧Manifold branch

170‧‧‧Manifold branch

172‧‧‧Public pipeline

Claims (18)

A flow cell includes: a plurality of inlets, each of the plurality of inlets is manufactured according to a size to receive a reagent flow of a corresponding reagent from a plurality of reagents into the flow cell; an outlet, which is manufactured according to the size The flow of reagents flows out of the flow cell; and a flow channel is located between each of the plurality of inlets and the outlet and is connected to each of the plurality of inlets and the outlet fluid Connected, the flow channel includes: a manifold section having a plurality of manifold branches in fluid communication with a common pipeline, each of the plurality of manifold branches is connected to a corresponding inlet of the plurality of inlets , And a detection section, which is in fluid communication with the common pipeline and the outlet, the detection section being operable to perform a plurality of different chemical reactions between the analyte located in the detection section and the multiple reagents Wherein the manifold branches of the manifold section have a first height and the detection section has a second height, wherein the first height is smaller than the second height, and wherein the manifold section has a sweep than the detection section The sweep volume is at least about 10 times smaller than the sweep volume. The flow cell according to claim 1, wherein the manifold section and the detection section of the flow channel are substantially planar. The flow cell according to claim 1, wherein the plurality of manifold branches are in fluid communication with the common pipeline through a plurality of forked joints, and each of the plurality of forked joints guides the flow of a corresponding reagent Passing the common pipeline and entering the detection section, at least one fork-shaped joint forms an acute angle between the corresponding branches of the manifold. The flow cell according to claim 1, wherein the flow channel includes a sweep volume and a geometric shape such that a flushing factor for achieving a flushing efficiency of at least about 99.95% concentration of the reagent located in the detection section is about 2.5 Or smaller. The flow cell according to claim 1, comprising: a top layer that defines the top surface of the flow channel; a bottom layer that defines the bottom surface of the flow channel; and an intermediate layer that defines the geometric shape of the flow channel. The flow cell according to claim 5, comprising a gap height defined by a distance between the bottom surface of the flow channel and the top surface of the flow channel, wherein the gap height is in the entire The flow channel is substantially constant and in the range of about 60 to 100 microns. The flow cell according to claim 1, wherein the flow channel includes a sweep volume and a geometric shape, so that a flushing factor for achieving a flushing efficiency of at least about 99% concentration of the reagent located in the detection section is about 2.0 or less. The flow cell according to claim 1, wherein: the plurality of reagents are from a plurality of corresponding reagent wells located in one of the cassette or the instrument, and one of the cassette or the instrument includes a plurality of reagents outside the flow cell Valves, each valve is located between a corresponding reagent hole in the plurality of corresponding reagent holes and a corresponding inlet in the plurality of inlets, so that each valve controls the plurality of corresponding reagent holes The reagent flow of the corresponding reagent well. An instrument with a flow cell includes: a plurality of reagent wells, each of the plurality of reagent wells can be operated to contain a reagent therein; a plurality of valves, each of the plurality of valves is connected to The corresponding reagent wells of the plurality of reagent wells are in fluid communication, and each valve of the plurality of valves is operable to control the flow of reagents from the corresponding one of the plurality of reagent wells; and The flow cell is operable to be fluidly coupled to the instrument, the flow cell comprising: a plurality of inlets, each inlet being in fluid communication with a corresponding one of the plurality of valves, Each inlet is sized to receive the corresponding test from the plurality of reagent wells The reagent flow of the reagent hole; the outlet, which is sized to allow the reagent flow to flow out of the flow cell; and the flow channel, which is located between each of the plurality of inlets and the outlet, and In fluid communication with each of the plurality of inlets and the outlet, the flow channel includes: a manifold section having a plurality of manifold branches in fluid communication with a common line, each branch connected to the plurality of One of the inlets, and a detection section, which is in fluid communication with the common pipeline and the outlet, and the detection section is operable to perform a plurality of differences between the analyte and multiple reagents located in the detection section The chemical reaction; wherein the manifold branches of the manifold section have a first height and the detection section has a second height, wherein the first height is smaller than the second height, and wherein the manifold section has a greater than The sweep volume of the segment is at least about 10 times smaller than the sweep volume. The apparatus according to claim 9, wherein the manifold section of the flow channel is the only common area in which the flow path of the reagent is pressed before flowing into the detection section of the flow channel The prescribed routes are transmitted together. The apparatus according to claim 9, wherein the manifold section and the detection section of the flow channel are substantially planar. The apparatus according to claim 9, wherein the manifold branch is in fluid communication with the common pipeline through a plurality of fork-shaped joints, and the fork-shaped joints guide each reagent flow through the common pipeline and enter the In the detection section, the fork-shaped joint only forms an acute angle between the branches of the manifold. The apparatus according to claim 9, wherein the flow channel includes a sweep volume and a geometric shape such that a flushing factor for achieving a flushing efficiency of at least about 99.95% concentration of the reagent located in the detection section is about 2.5 or smaller. The apparatus according to claim 11, comprising: a top layer which defines the top surface of the flow channel; a bottom layer which defines the bottom surface of the flow channel; an intermediate layer which defines the geometric shape of the flow channel; and a gap A height, which is defined by the distance between the bottom surface of the flow channel and the top surface of the flow channel, wherein the gap height is substantially constant throughout the flow channel. The apparatus according to claim 9, wherein the flow cell is positioned in the apparatus. A method of using a flow cell includes: connecting the flow cell to an instrument, the flow cell including a plurality of inlets, outlets, and flow channels in fluid communication between the plurality of inlets and the outlet, the The flow channel includes a manifold section and a detection section, wherein the manifold branches of the manifold section have a first height and the detection section has a second height, wherein the first height is smaller than the second height, and wherein the manifold The pipe section has a sweep volume that is at least about 10 times smaller than the sweep volume of the detection section; the first valve among the multiple valves of the instrument is operated to select the first reagent among the multiple reagents, and each reagent is located in the box or In the reagent well of one of the instruments; pump the first reagent through the first inlet of the plurality of inlets of the flow cell and through the flow channel of the flow cell; The first chemical reaction between the analyte in the detection section of the flow channel and the first reagent, wherein after the first chemical reaction is completed, at least some of the first reagent remains in the Remaining reagents in the flow channel; operating subsequent valves of the plurality of valves to select subsequent reagents of the plurality of reagents; and pumping the subsequent reagents through the subsequent inlets of the plurality of inlets and through the flow aisle, The remaining reagent is flushed out from the flow channel, so that the reagent with a concentration of at least about 99.95% in the detection section is the subsequent reagent using the total flush volume of the subsequent reagent, and the total flush The volume is equal to or less than about 2.5 times the sweep volume of the flow channel. The method according to claim 16, comprising: after a concentration of at least about 99.95% of the subsequent reagent in the detection section is reached, executing the subsequent reagent and the subsequent reagent in the detection section A subsequent chemical reaction between the analytes, wherein after the subsequent chemical reaction is completed, at least some of the subsequent reagent is left in the flow channel as a second remaining reagent. The method according to claim 17, comprising repeating the following actions: operating the subsequent valve, pumping the subsequent reagent, and performing a subsequent chemical reaction.

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